Luminol Tests Negative For Blood

Professor Francesco Vinci Luminol Report Amanda Knox case
Page 61 of Professor Vinci’s Report shows the print the Polizia Scientifica said was ‘compatible’ with Raffaele Sollecito. Clearly it’s not a match. In fact, it’s shorter by 3cm!

The luminol tests were done on December 18, 2007 after the whole flat had been turned upside-down during multiple undocumented inspections and revealed the presence of six “footprints” testing positive for DNA and three more that tested negative for DNA. All of the prints and blobs tested negative for blood with the tetramethylbenzidine [TMB] test. Prosecution forensics expert, Patrizia Stefanoni, was asked about TMB [concerning tests on the knife] at the pre-trial in October 2008 and said a negative result gives certainty it’s not blood.[1] Stefanoni failed to disclose the negative TMB results when she testified at both the pre-trial and trial, nor were they recorded in the Polizia Scientifica RTIGF Report [PDF pages 223-229, original 218-224]. It was only after the court ordered a partial discovery data dump on July 30, 2009 that the defense became aware.

On September 26, 2009 Knox defense consultant Dr Sarah Gino returned to court and revealed everything tested negative for blood and the prosecution’s insinuations and media leaks about bloody footprints and clean-ups was bogus. She further revealed there was still missing data and the mixed trace in Filomena Romanelli’s room was low copy number [LCN] single amplification. ie not a valid result and possibly from contamination.[2] For some unknown reason, the Scientific Police didn’t photograph the luminol in Romanelli’s room or if they did, it wasn’t disclosed. Also strangely, it’s not listed on the 2008 Scientific Police evidence map.

Professor Vinci Report: Key failures in the work of the Scientific Police

  1. Preliminary study of the feet was not carried out by the Scientific Police for morphological and anatomical characteristics, static and dynamic tests, x-ray exams. According to Robbins et al, “It is not possible to analyze a footprint without having any anatomical knowledge of the structure of a foot”;
  2. The prints left by the feet of the defendants were taken using a method appropriate for other types of investigation; in any case no print was taken “dynamically”, i.e. through a prolonged walk of at least 6 meters, as is widely recommended in literature: it does not appear that this was done in the case in question;
  3. Literature recommends the use of a soluble paint in water which has an ideal degree of viscosity for the collection of footprints. This was not done by the Scientific Police. Instead it is not recommended to use the method of “imprinting” in order to obtain a footprint. In the example the prints are in fact too dense for the requirement;
  4. During the collection of the defendant’s footprints for analysis it is recommended to use a method to get their attention so as to render their walking as natural as possible. This was not done by the Scientific Police.
  5. The Scientific Police did not carry out a preliminary evaluation of the prints;
  6. Direct examination of the samples: The Scientific Police did not carry out an evaluation of: the composition of the material, its characteristics, permeability of the warp and weft, observations with alternative lighting [Crimescope], chromatic reactions with various illuminations;
  7. Dr. Rinaldi declared to have only examined photographs;
  8. Choice of system of measurement: The Scientific Police did not indicate their method of measurement.

Other Investigation Failures: Reference DNA and footprints samples

  1. The only DNA reference samples taken were from the defendants, Lumumba and the victim;
  2. For the footprints: Only the three accused gave a reference footprint. Not even the footprint of the victim was taken, nor the other residents of the cottage;
  3. Fluorescent metric ribbons weren’t used to measure the prints;
  4. The photographs weren’t taken in perspective but at an angle.

Luminol Photos

The high resolution crime scene photos below are dsc 146 & 147 taken outside Meredith Kercher’s bedroom door. The photos can be clicked to enlarge or accessed in the gallery on this site. Firstly, what you see below tested negative for blood. The prosecution tried to say it’s cleaned up blood. Asides from testing negative for blood, there’s no swirling pattern to indicate whatever it is has been cleaned up at all and there’s gaps between where it’s reacting. A liquid being cleaned up would leave a continuous joined together pattern not dots all over the tiles.

The photos are highly suspicious because if you look closely, you can see the luminol lighting up on top of the investigators foot in the bottom left of the photo, and on a little pebble and even on top of the police measuring ruler. How is that possible? You can also see whatever it is has seeped into the grouting. People are free to speculate about what it is and how and when it got there. It’s important to note the burden is on the prosecution to prove it’s blood not on the defense to explain what it is. At the trials the prosecution reversed the burden of proof. Judge Massei ignored the negative TMB results and said because it’s near where a lot of blood was in Meredith’s room, it must be blood which is clearly absurd reasoning.

 Meredith Kercher murder luminol photo 147  Meredith Kercher murder luminol photo 146  enhanced luminol photo amanda knox case
High resolution crime scene photo dsc 147
High resolution crime scene photo dsc 146
Enhanced photo of dsc 146 
luminol on top of the crime scene investigators booties luminol on a pebble in the amanda knox cottage luminol on top of the police measuring ruler meredith kercher murder
You can see it lighting up on top of the CSI investigators foot
Here you can see the luminol lighting up on top of a little pebble
And even on top of the police ruler. How?

Luminol Lab Work

  1. The luminol samples from Sollecito’s place were all extracted as “Saliva,” except 101 [Uomo 6] per SAL. Note – that if “Saliva” is listed, the “TMB” field does not appear in the SAL. Also, there are no quantification records for these.
  2. Rudy Guede’s place — they only used luminol on one spot? Seems unlikely, so a number of items that are possible luminol hits are listed. No SAL for the samples from Guede’s and no quantification records, either. 12 out of 13 STRs missing. What are they hiding?
  3. Via della Pergola 7 — Sarah Gino is using an LCN definition of < 200 pg/uL, although Stefanoni ran some samples with way less DNA than these samples. Note the re-runs for Reps. 176, 178, 179. Every print/blob other than Rep. 180 is LCN.

Amanda Knox case

Testimony Excerpts

Murder of Meredith Kercher footprint map from the Amanda Knox cottage
This is the only luminol footprint map produced by the prosecution’s experts. It’s from page 15 of the Rinaldi-Boemia report.

Patrizia Stefanoni Testimony Pre-trial October 4, 2008 p177 [A negative TMB result means it’s not blood]

Judge:  Ok! And here there is a degree of sensitivity?
Answer: It is very sensitive, now I do not know how to say it to him, however, in common practice …
Judge: There also cites false positives of the series …
Answer: Yes, in the sense that it does not distinguish whether it is human or animal blood, for example.
Judge: However where the result is negative I’m given to understand that it’s almost certain that it is not [blood]?
Answer: Yes, it’s not blood, it is not, yes.

Sollecito Defense Consultant Professor Tagliabracci July 18, 2009 p71

Answer: […]tetramethylbenzidine is a very sensitive diagnosis that can highlight up to five red blood cells. So that a negative result in short leaves no room for doubt

Patrizia Stefanoni Testimony May 23, 2009 p144 [Only 5 reference profiles were collected]

Question: Just to clarify here before answering the questions of my colleague has said that you had a profile … four genetic profiles.
Answer: By reference, yes.
Question: In fact there were five.
Answer: And the victim’s.
Question: Of course the victim’s, then we say that the genetic profiles were five in total, the two defendants, the victim, Lumumba, and Rudy?
Answer: Yes.
Question: And nothing else?
Answer: No

Knox Defense Consultant Professor Carlo Torre page 6 [Torre testified before the defense discovered everything tested negative for blood]

Luminol print that doesn't match Amanda Knox
One of the luminol prints attributed to Amanda Knox. The 2nd toe is significantly shorter than Knox’s.

A couple words only on luminol traces, now we know that importance was given to a few traces, for example the one in Amanda Knox’s room for which a blood testwas not done, my colleague DNA expert Gino will talk about it after, and trace 7, these are the ones that concern us more directly from which a specific test for blood was not done and from which no genetic profile was extracted. Now the prints are these, in consultation with Rinaldi and Boemia one says “prints impressed by depositing blood” but how can you say that those prints were made in blood, you can say those prints are luminescent in reaction to luminol, we all know that the prints that we… there are various other materials that become luminescent with luminol, Sarah spoke to me about a really funny case maybe she wanted to speak about it, but there was a case in our area, of traces that glowed with luminol that created problems in the course of the investigation [inc] of blood and not blood, then it was clarified to be… because they were small because if not maybe one wouldn’t be able to understand that they were the traces of the child that had swam in the pool and got out with chlorine-covered feet, leaving this whole path, now I imagine if those traces weren’t made by a child how many innumerable measurements the Polizia Scientifica would have done, and they probably would have written that they were deposits of blood. I add that, holy heaven no… in trace 1 DNA was taken, in trace 7 nothing, now with this luminescence it’s difficult to think that DNA would not have been extracted, it’s a strong luminescence, first it would be appropriate perhaps to give a specific diagnosis of blood but, my colleague will speak about these things.

Now we have a plantar print assigned to Knox and luminol trace 1, I think the experts are Boemia and Rinaldi, they speak in this case about probable identity, now be careful because first of all the literature advises always to not just take one print only but different positions. I don’t know if in this case it might have been done because the print can change according to the behavior of the foot, from an external pressure, internal and so on but I am not very expert on prints and I say it right away, yet the measurements I know the fact that they were consistently incorrectly measured was discussed I want to let you know that there exists on the market these luminescent glowing strips that are made for the sole purpose of placing next to luminol traces to be photographed in order to have a precise metric point of reference, and then not work from photographs. It’s longer, it’s shorter, the photo is crooked, for which it is difficult to reach a measurement of any interest but above all I ask myself this – These people made a sea of calculations, I’ve read a bit of the literature on these things but here I hadn’t… It’s not a question of reading the literature, it’s clear that this foot of Amanda’s has the second toe let’s call it index of the foot to make ourselves more understandable, longer than the big toe. In the print the second toe is frankly shorter, in the luminol, both Amanda’s foot and its ink footprint have a long second toe, that print there with the Luminol has a short second toe I’d say there’s no doubt about it. I’m not particularly an expert on prints but if there’s one thing that jumps out when we know that the majority of people have a second toe longer than their big toe however a good number people have a shorter one. I believe that it would not have been a bad idea to take the prints of the other young people that came to the house and that potentially walked on the floors with bare feet perhaps coming out of the shower that maybe had been washed with chlorine to clean it, that could have left prints that were Luminol positive anywhere.

I don’t mean to say but unfortunately I have no precise image, the only other foot that I could see was that of the victim. Well, there’s no image in which you can see it clearly but if I must say the foot of the victim, it would be good for whoever has images of the victim’s foot available to study them, it seems that she has a shorter second toe. You can’t tell because here it is slightly flexed, this to say that the same victim could easily the day before, two days before, have taken a shower and have walked across that floor and left a luminol positive print. I’m not saying everywhere but almost. Probable identity, even here the same things are true even for the second print you see clearly that here there’s no trace of the long second toe. So, speaking this probable identity that these people talk about, there exists… it’s accepted by ENSI, it is described among other things in an Italian text as recent as 2009 and in which I think even Intimi who I think is the director of the Scientific Police participated. I know little of police work I don’t know the degrees, I don’t know how they are called, anyway identification, this makes reference to shoes be careful however… there does not exist an analogous criteria on the subject of footprints however the general concept I’d say is that there is identity when, upon comparison, the same general characteristics and particulars present themselves.

Probable identification, this is what our friends in the Scientific Police say but be careful to speak about probable identification, there’s a clear correspondence of design and form in the absence of significant differences more than a coincidence of the general characteristics or of any markers of wear, in addition at least one of the following criteria must be satisfied, are there observable markers, what is a marker? In forensics you study characteristics and markers, characteristics are blue eyes, brown hair, and they make up an anatomic inventory of each human they are the characteristics and therefore can have a certain value in identifying a person, I can say that that person with blond hair and blue eyes, the markers on the other hand are something anomalous, a tattoo, an amputation of a finger, a wound, a particular surgical intervention, so for these prints to reach a probable… to have an identity they must be the same, to have a probable identity they must at least have either some attributable markers in terms of their fabrication so I’m talking about shoes or casual markers for example I walked once on some nails and they poked holes in me ,or there’s a clear agreement in the specific markers that are possibly not definable and so on, anyway that’s all. Be careful again concerning… I almost finished, regarding the luminol the literature insists on the fact that luminol can easily diffuse and can alter the look of foot prints.

Amanda Knox case: Rep176
Rep 176 – Low Copy Number, single amplification, TMB negative and strangely no photograph of the luminol reacting was taken.

Knox Defense Consultant Dr Sarah Gino September 26, 2009 page 6

Consultant. – I would like to proceed to my observations at last: the traces we discussed last time, luminol-positive traces. What emerges from the documents disclosed in July 2009? These luminol-positive traces were named exhibits 176, 177, detected in Romanelli’s room, traces 178, 179, 180 detected in Knox’s room, and exhibits 181, 182, 183, and 184 found in the corridor outside the victim’s room. These tracks, let’s recall, are those tracks that were brought into evidence by the spraying of luminol. Analyzing these SAL cards, we learn – in contrast to what the technical report of the Scientific Police represents, and to what has been claimed in the courtroom – that not only the luminol tests were performed but these traces were also subjected to a generic blood tests with {74} tetramethylbenzidine.

Tetramethylbenzidine is the test we normally use in the lab to find out if a trace is possibly blood or not; it is a very sensitive method; I believe that Professor Tagliabracci during his last round of testimony stressed that repeatedly; however it is not specific because we have seen there exist false positives with this… with tetramethylbenzidine, so something that gives a positive result is not blood in reality. This data, however, is new; only now that we have the SAL cards do we know that a second test was performed; and what did that test produce? It produced negative results for the exhibits for which it was possible to obtain a genetic profile, that is 176, 177, 178, 179, 180, 183, while for exhibit 183, the other three exhibits… sorry, exhibits 181, 182 and 184 gave an “uninterpretable” result.

At this point it seems to me justified to question whether these luminol-positive traces, which have been much discussed, can still be considered traces of a hematic nature; further, analyzing the quantification data we see that the quantity of DNA recovered from most of these traces should be compatible with LCN DNA, so with DNA present in low amounts; also in this case it is necessary to question whether the amplification was repeated or not to be able to consider the result obtained scientifically valid; these are my remarks on the documents delivered to us.

Counsel Ghirga. – I…
President. – Please, counsel.
Counsel Ghirga. – Regarding tetramethylbenzidine, the chemical test for blood, on exhibit 176, as you said, negative; on exhibit 170, positive, relatively (inaudible) and 170 of the technical report already in the records from June 2008, you referred to these samples when you said… {75}
Consultant. – Yes.
Counsel Ghirga. – And this negative, how… where it says negative, what should we think, is it negative for blood?
Consultant. – When it is negative, because I am running a test on a substance which I assume is blood because of the luminescence, then it is obvious tha`t I am looking for presence of blood, if it comes back negative, this presence of blood cannot possibly be [non può assolutamenta essere] established.

[….]

Counsel Del Grosso. – So, Dr. Gino, referring to what you have said, the luminol-positive traces – we have talked of the technical report – what information are we getting?
Consultant. – From the technical report, we only learn that these traces are those which at the site of the investigation had a positive result in what is called a generic diagnostic, that is positive in a generic test made by luminol. {77}
Counsel Del Grosso. – I recall there is a little square with the words: “generic diagnostic”; there is an X to say “positive according to the diagnostic.”

Consultant. – There’s a positive V but in the little square nearby it’s written down; anyway, now I’m saying this in my own words, the concept is this: made in the course of the investigation with luminol – but we have seen for other traces, for example the trace where it said…
Counsel Del Grosso. – Closer to the microphone, please.
Consultant. – Where it said, I think it’s 170, the square [il quadrati? either “i quadrati” or “il quadrato”]… it is written: “test made with generic diagnostic tetramethylbenzidine ” and it’s a positive result.
Counsel Del Grosso. – However, by checking the SALs, that is only by checking these documents we have found out that a specific test for blood was administered.
Consultant. That’s a test which also…

Amanda Knox case: Rep 177 – Low Copy Number, single amplification, TMB negative and strangely no photograph was taken of the luminol reacting.
Rep 177 – Low Copy Number, single amplification, TMB negative and strangely no photograph was taken of the luminol reacting.

Counsel Del Grosso. – This is the test whose name I cannot master…
Consultant. – Testing with tetramethylbenzidine, it’s a test like luminol – also presumptive because he does not tell us whether that is really blood. A positive result tells me it could be blood, it could be blood of a dog, a horse and a human. To find out if this is truly blood, which contains hemoglobin, it is possible to run a test, e. g. with immunochromatography to learn if that is human hemoglobin, or other types of analysis instead which can indicate to me the presence of hemoglobin but do not tell me whose blood it is, therefore this test tells me that blood could be present.

Counsel Del Grosso. – But the result was negative?
Consultant. – But the result was negative.
Counsel Del Grosso. – Thank you.
President. – There are no further questions, for the counter-examination only on the last aspect obviously because it was been {78} exhausted…

[….]

Counsel Del Grosso. – Page 138, hearing March 23, 2009. Referring to the luminol traces, I’d like to seek clarification. You were able to ascertain from the documents produced that one is dealing in this case with LCN?
Consultant. – Yes, as I have mentioned before, it follows from the quantification {90} that the quantity of DNA present in this trace, if I may consult the documents…
President. – If they are at your disposal, it is authorized.
Consultant. – I’m going to do it… I’m going to give an actual example, that is we won’t be talking in the abstract but, for example, of sample 176… the quantity of DNA is 0.04, we are talking of nanograms/microliter so it is means 40 picograms/microliter so we are well below the minimum threshold of which we have previously spoken, so some of these findings [exhibits], I would say practically all of them with the exception perhaps of 180, can be considered LCN.
Counsel Del Grosso. – Oh, and…
Consultant. – However as for this matter, we do not know if the guidelines were followed, if they were re-amplified, if it was verified that the same type of profile turned up more than once.

Counsel Del Grosso. – Excuse me if this question is irrelevant, between a datum, a quantity below 10 picograms and a quantity of several hundred picograms, for you geneticists dealing with such quantities every day, is there a difference that you can determine to be material, low, we may fail to understand it in quantitative terms perhaps but below 10 picograms, what can an amount be equal to?
Consultant. – It’s a minuscule amount, it’s an amount that we should anyway always bear in mind that even if I amplify – because I’ve decided that I’m going to take everything I have and amplify everything I have even if it’s below the amount normally used – [and] it’s an amount that gives rise to a genetic profile, I must always put this question to myself, whether this genetic profile was not generated from something else, so it is not derived from a contamination [event]; there’s a difference between talking of hundreds and of a few tens of {91} picograms just as there’s a difference between talking of one nanogram and 100 picograms: one nanogram is a good amount of DNA that lets me work in full confidence; 100 picograms, 200 picograms, below 200 picograms, as I was made to point at the last hearing [by] the consultant of the prosecutor, but I would say that below 100 picograms one should take care, that is I must pose the question, if I get a result, how to interpret it.

Counsel Del Grosso. – These guidelines which have been cited several times in these evidence hearings, which prescribe these precautions, a double amplification and whatever we have referred to, serve and are necessary to avoid and stem the risk of contamination which is…
Consultant. – Yes, certainly, because…
Counsel Del Grosso. – Tell us why?
Consultant. – It’s that… naturally, all that can help us because obviously repeating genetic profiles, rerunning amplifications, thus obtaining genetic profiles from the same extract of DNA, we can consider only those alleles, i.e. peaks, good and valid which repeat themselves every time we run an amplification. In contrast, we must consider the peaks which do not repeat themselves to be spurious, and the spurious peaks can derive from stutter[s], as has been surely said, from the stutters, thus this erroneous amplification of the allele, or rather, of other DNA which travels around – it is said that DNA flies, DNA flies not [because] it has wings but in the sense that all of us shed DNA and it’s true that when… in a laboratory where they amplify hundreds upon hundreds of samples every year some trace of amplified DNA can be present in the tools, can be present on the pipette I use, can be present in the hood under which I prepare my reactions; this is a reality that must be taken into consideration: one cannot eliminate that {92} contamination; one cannot say: “My laboratory is the best of all because we have never had contamination” – that is simply not true [as] cases of contamination exist; it is necessary to take this into account, above all these instances of contamination that happen when one works in extreme conditions, that is with a few…

Counsel Del Grosso. – But with these infinitesimal amounts, is the risk of contamination greater?
Consultant. – It’s not that the risk of contamination should be greater; it is more likely [lit. easier] that I should find the contamination, hence the risk of contamination is the same.
Counsel Del Grosso. – Certainly, but is it more…
Consultant. – When from the sample… however it is easier to find that out because we may have zero DNA of my extract with a tiny amount of that contaminant, it is obvious that I should know that, while I have, instead, a great amount of DNA of my extract and that tiny amount of the contaminant, it’s an overwhelming amount of DNA and of the extract and therefore I am ignorant of that contamination.

[….]

Counsel Del Grosso. – Regarding trace 177, one of the luminol-positive ones that we said was LCN, is there a way to retrieve other genetic profiles in addition to those already found?
Consultant. – Yes, I stressed that at the last hearing, I brought forward… exhibit 177 as an example, where I pointed out that in addition to the profiles… that is, other that the peaks related to the victim and those attributed to {93} Amanda Knox there were other peaks which were not taken into consideration although some were higher than 50 RFU, which we have always talked about.
Counsel Del Grosso. – Now as regards only the luminol traces, I wanted [to ask] if you remember, whether during the oral testimony before the GUP by Dr. Stefanoni – page 58 and further – she spoke of only having run a generic test on these traces, the luminol positive [test], but not of having performed other tests, which emerged today…
Consultant. – Yes, in fact it emerged that the tetramethylbenzidine test was negative for most {94} of the traces and was non-interpretable for two.
Counsel Del Grosso. – Thank you, no more questions.
President. – Good, as there are no more question, Dr. Gino is dismissed, the report shall be obtained together with the other documents produced by the defense of Amanda Knox.

Patrizia Stefanoni Testimony May 22, 2009 page 52

Amanda Knox case: Luminol
Patrizia Stefanoni May 22, 2009. “because there are, indeed, so many possibilities of having false positives” “this is why it is easy to have false positives”. This was ignored by the courts.

Answer: No, in fact, as is reported later, the positive [response] to the test does not indicate with certainty the presence of human blood, and nor does it indicate with certainty the presence of blood, so if I have animal blood, such as fish blood, blood of meat that is commonly used, commonly handled, commonly bought in households, I nonetheless can have in those points where maybe there had been meat, fish, thus animal blood, I can nonetheless have a positive [reaction] to Luminol. I can have, however, a Luminol luminosity also in other circumstances, when for example I have a surface where there has not been blood placed against/on it, thus it has not had contact with blood, but maybe there’s rust that I cannot see with the naked eye, [or] there’s, I don’t know, fruit juice – some fruit juices give positive [reactions] to Luminol – [or] there’s also been grass, so also plant chlorophyll gives positive [reaction] to Luminol. All these situations are defined, indeed, as false positives. With the Luminol, I cannot distinguish/differentiate them, thus I cannot grasp whether this is a false positive or a true positive. I must however… how can I say, take note of them, document them, if I can, from a photographic point of view and attempt to analyse them, so a negative result to the DNA test on traces found with Luminol does not necessarily mean that there [in that point] there was no human blood: it might be that there [in that point] there was in fact no blood, so there was no human blood, maybe it was grass, [or] it was chicken blood, and so on. So I, with genetic analysis, can say with certainty that there was blood, but its absence, thus the absence of a DNA profile [53], does not allow me to establish much: it does not allow me to say if there was no DNA because there was too little, thus there was no blood and therefore there was DNA at all, or else there was something else that was not blood, there was maybe grass, grass residues that were trampled on. The luminescence that is given off is a very intense colour, very particular because it is blue, azure, and so it is visible for a few seconds, then declines/fades. It is as though that luminosity turns itself off and thus we can no longer see it: the reaction is spent/depleted.

These are examples that regard this case: they are examples in fact of positive Luminol traces, and as you can see, these are plantar prints, clearly, in short, of bare feet, after the luminescence with Luminol. And here this piece of flooring is shown/reported that corresponds to this, not to this, but to this one here, where as can be seen on two of these intense fluoresences that we have here to the naked eye, with the light [of day] absolutely no trace is visible, no print, so no haematic trace nor any other type [of trace] is visible to the naked eye, whereas instead after Luminol one has this fluorescence which, however, as you can see, nonetheless has a background, because there are, indeed, so many possibilities of having false positives, so there could be a basis/bottom of … I don’t know, of earth, of rust, of something that mimics, but in a more faint way, thus a more feeble luminosity, less intense, let’s say, [than] the luminosity that on the contrary is given off in correspondence to the points that we then choose to sample. I have not told you that Luminol reacts … in effect it uses [54] as one of the ingredients of the reaction the iron contained in haemoglobin. This is why it is easy to have false positives, because rust also has iron, also chlorophyll does not have iron, but it has a molecule that is very similar to haemoglobin that contains iron, and anyway there is another atom that, let’s say, replaces the iron in effecting this type of reaction, and this [atom] is magnesium. So in effect, iron can be shall we say, indeed, indicative of the presence of haemoglobin, but iron is not the only [atom] that reacts with Luminol: other chemical elements also react, also other metals, and then iron is not the sole constituent … that is, it is not contained in a specific manner in haemoglobin, but it is an atom [which is] contained in a great many molecules, both organic and inorganic.

Luminol At Raffaele’s Apartment

Patrizia Stefanoni's Report Page #68
Patrizia Stefanoni’s Report Page #68

Patrizia Stefanoni's Report Page #69
Patrizia Stefanoni’s Report Page #69

Patrizia Stefanoni's Report Page #70
Patrizia Stefanoni’s Report Page #70
Overall Analytical Results
Apartment in use by SOLLECITO Raffaele
Technical inspection performed on November 13, 2007
EXHIBITS and TRACES
Kitchen – Entrance
TRACE TYPOLOGY
GENETIC RESULT
Blue Sponge
Exh.90 / A
hematic substance: negative
SOLLECITO
Siphon under the sink
Exh. 91/A/B
hematic substance: negative
NEGATIVE
EXHIBITS and TRACES
Bedroom
TRACE TYPOLOGY
Highlighted by
luminol
GENETIC
RESULT
Outside door handle
Exh. 92 / A
presumed blood substance
NEGATIVE
Floor sample Exh.93 / A
presumed blood substance
SOLLECITO + KNOX (partial)
Floor sample Exh.94 / A
presumed blood substance
NEGATIVE
Overall Analytical Results
Apartment in use by SOLLECITO Raffaele
Technical inspection performed on November 13, 2007
EXHIBITS and TRACES
Bathroom
TRACE TYPOLOGY
Highlighted by luminol
GENETIC RESULT
Floor sample
Exh.95 / A
presumed blood substance
SOLLECITO + KNOX
Floor sample
Exh.96 / A
presumed blood substance
KNOX
Underside shower bathmat
Exh.97 / A
presumed blood substance
NEGATIVE
shower base
Exh.98 / A
presumed blood substance
NEGATIVE
Interior door handle
Exh.99 / A
presumed blood substance
NEGATIVE
Outside door handle
Exh.100 / A
presumed blood substance
NEGATIVE
Overall Analytical Results
Apartment in use by SOLLECITO Raffaele
Technical inspection performed on November 13, 2007
EXHIBITS and TRACES
Kitchen – Entrance
TRACE TYPOLOGY
Highlighted by luminol
GENETIC RESULT
Bathmat upper surface
Exh.101 / A
presumed blood substance
Male individual MAN #6
Floor sample
Exh.102 / A
presumed blood substance
NEGATIVE
Floor sample
Exh.103 / A
presumed blood substance
NEGATIVE
Floor sample
Exh.104 / A
presumed blood substance
NEGATIVE
Floor sample
Exh.105 / A
presumed blood substance
NEGATIVE

There were a total of fourteen luminol-positive areas in Raffaele’s apartment. Ten did not yield a DNA profile; one had Amanda’s DNA profile; two had a mixed profile from Amanda and Raffaele, and one had the DNA profile from an unknown male. Three came from bedroom or bathroom door handles; five came from the kitchen, two from the bathroom, and two from the bedroom. The Massei report stated [p. 194, English translation], “The November 13 inspection in the apartment used by Raffaele Sollecito did not yield any significant results.”

The lack of any DNA in ten samples is suggestive that they were not made in blood. The lack of Meredith’s DNA profile in the remaining samples means that it is unlikely that any of those four stains was made in Meredith’s blood [from a putative clean-up]. The presence of a DNA profile from an unknown male highlights an important issue: the DNA profiles could be unrelated to the substance causing the luminol reaction. If areas near the four DNA-positive, luminol-positive stains had been tested and found to lack DNA [these are called substrate controls], then there would be some reason to associate the DNA with the stain, but there were no substrate controls performed. Moreover there is no record of confirmatory [antibody-based] blood testing; therefore, there is no reason to conclude that any of the luminol-positive areas are blood. Among the many substances giving false positives with luminol are animal blood, rust, and draino. The large number of luminol-positive areas is not surprising, given that it is a presumptive test for blood, not a confirmatory test, and Sarah Gino’s testimony indicated that luminol is prone to false positive reactions.

Court Rulings

Italian Supreme Court

The 2013 Italian Supreme Court ruling is Kafkaesque regarding the luminol and completely ignores the 250 other substances it reacts with and the negative TMB results. This excerpt is from the Analysis of Prints section of the Chieffi Report:

The logical rigour adopted in a further passage of the Court’s reasoning appears to be just as inadequate. This relates to the confirmed presence of traces revealed with luminol (as they could not be seen with the naked eye) which yielded Knox’s profile and the mixed profile of Knox and Kercher, and which were found in Romanelli’s room, in Knox’s room, and in the corridor. These traces could not be attributed to prints left on other occasions, as the court of appeal implausibly held, since luminol detects traces of blood, and it is not really conceivable that Knox’s feet were stained with the victim’s blood on previous occasions. As the petitioning party noted, the odd coincidence of Knox’s DNA in all the traces mixed with the victim’s blood was not accounted for, whereas the theory proposed in the first instance ruling far more plausibly gave importance to the mixed nature of the traces (including those found in the small bathroom). It concluded with adequate logic that Knox had cleaned her bare feet of the victim’s blood but that residual traces remained, and that she went into her own and Romanelli’s rooms by way of the corridor to carry out the staging considered in the initial reconstruction of the crime to have taken place.

Nencini Report

The luminol stains testing negative for blood are not mentioned anywhere in the entire motivation report.

Hellmann Report

Prints Revealed by Luminol With Useful Biological Profile

It was during the second inspection at the cottage at Via Della Pergola 7, carried out by ERT [Esperti Ricerca Tracce – Trace Search Experts] Scientific Police on the date of 12/18/2007, that aspersion with luminol was performed of the floor of the corridor, the kitchen/living-room, the bedrooms of Amanda Knox and Filomena Romanelli, and the larger bathroom. Chief Inspector Ippolito took photographs of the traces of bare footprints – detected only in the corridor/living room and in Amanda Knox’s bedroom. While taking the pictures he did not use fluorescent metric ribbons, which would have been useful for the subsequent measurement of the photographed footprints (ruling page 369).

The first-level ruling reports on these traces [footprints] (Exhibits 176 to 184) on pp. 303 et seq., specifically concerning those testing positive on genetic investigations, and also on pp. 370-373 on those which, in the absence of genetic findings, were submitted to morphological and dimensional examinations. The prints are of bare feet, detected in Romanelli’s room (176 and 177), in Knox’s room (178, 179, 180), in the corridor (184, rectius [Latin: “more correctly”] 183). According to the indications in the SAL [Stato Avanzamento Lavori, “State of Work Progress”] files of the genetic lab of the Scientific Police, the generic test for blood was performed on these footprints, which gave a negative response.

The genetic investigations, conducted by Dr. Stefanoni, biologist of the Scientific Police, gave the following results: 176 trace of Meredith; 177 mixed trace of Meredith and Amanda; 178, 179, 180, biological profile of Amanda, 184 (rectius 183) mixed genetic profile of Meredith and Amanda. During the first-level trial, Dr. Sarah Gino, consultant for the defense, stressed that the quantity was compatible with low-quantity DNA (low copy number), that the analysis had not been repeated to validate the result and that peaks were present, which had not been considered, that could show the presence of other contributors. She also posed the problem of contamination — of non-authenticity of the traces — and consequent irrelevance of the traces themselves.

The ruling, though noting that the number of substances reacting to luminol is rather large, gave arguments [purporting] to rule out (p. 304) that the material covering the floor of the house on Via Della Pergola had such a quality. It assessed as improbable the possibilities suggested regarding specific substances, stating (p. 305): “one would have to hypothesize that one of those substances (certain vegetables, fruit juice, rust, bleach…) had been on the floor where the luminol test was performed, and, present on the date of December 18…would have been affected by some biological trace that placed itself on one of those luminol positive substances, a biological trace originating from Amanda and in two cases also from Meredith”.

Specifically concerning bleach, though noting the likelihood that such a product had been aspersed everywhere in the house during ordinary cleaning, the ruling concluded that, in actuality, “it cannot be known when and by whom such a pervasive and extensive cleaning involving the various rooms was carried out. Moreover, nobody entering the house claimed to have noticed the smell of bleach…Furthermore, had bleach been applied in the whole house, in a cleaning operation involving the various rooms, many more luminol positive traces than those found should have been detected”.

The ruling thus concluded that the footprints had necessarily been left in Meredith’s blood, trodden on by Knox in the murder room and then transported by her into the other parts of the house. The belief of the first-level Court has been contested by Knox’s defense and faces insurmountable contradictions, both logical and factual. First and foremost, the certain, true fact is that the generic blood test gave a negative result. According to the [first] Court this happened because of the scarcity of the available biological material, but the consultant for the defense, Professor Tagliabracci, specified, without being refuted (hearing of July 18 2009, p. 174), that the tetramethylbenzedine (TMB) test is very sensitive, so much as to give a positive result even with only five red blood cells present. Dr. Stefanoni herself, moreover, clarified (preliminary hearing of October 4 2008) that, while a positive test result could be deceptive due to reactivity of the chemical [evidenziatore] with other substances, a negative result gives certainty that no blood is present.

Secondly, given that there are bleaches without smell or even perfumed – in any case luminol reacts to other cleaners too – it is not impossible to presume who may have performed the cleanings of the house. Four girls lived together in the apartment, and it appears reasonable to surmise that they, possibly in turns, accomplished the task [provvedessero alla bisogna], the more so since the house was visited by other people as well, friends or boyfriends as they might be, and so was even more subject to getting dirty. The limited number of footprints detected can be explained by treading at different times and by the use of bleach on points specifically dirty. After all, doubts similar to those raised in the ruling could be brought up even supposing that the traces are of blood: why only in those few points, moreover not consecutive and instead spread in various rooms?

The ruling arrives at an unlikely explanation (p. 306): Amanda “with bare feet, washed of Meredith’s blood, but under which there should still have remained blood residue, went to her room, to Romanelli’s room and passed by the corridor and in some places of the room, where she walked, she left the traces that were detected”. The paradox of an inaccurate foot-washing aside, what remains without an explanation is the fact that the traces were not left one after the other starting from the door of the small bathroom – moreover, about this issue see below – but instead just one in the corridor, at the wall dividing Meredith’s room from Amanda’s room (hence a few steps away from the bathroom); two even further away in Romanelli’s room; three in Amanda’s room.

[This is an] even more valid consideration in light of the serious contradiction in the ruling, where (pp. 408 and 409) it assumes that, instead, the footprints were left by Amanda with blood soaked feet and that she would have washed them only later.

Conscious of the weakness of its theory, the ruling takes for granted (p. 413) that a clean-up was carried out. It reaches such certainty considering that the foot which left the print on the mat could have arrived there only through steps that should have left other, even more obvious prints on the floor. Even neglecting to consider that, as has been seen, the footprint on the mat has been attributed to Raffaele Sollecito and not to Amanda, the explanation for the incongruity highlighted by the Court can be found in the statements of Amanda herself, who said that she took a shower the following morning and that she went back to her room dragging the mat with her bare wet feet, which was then put back in its place. Confirmation of this has been given by Professor Vinci, who examined and photographed the mat, revealing that it showed bloodstains on the bottom, which did not correspond to those on the upper side (p.37 of his report).

Beside which, the occurrence of a clean-up is negated by the sheer number of traces found in the house. To overcome this objection, the first-level Court goes as far as stating that during the clean-up the shoeprints were purposefully spared, in order to direct suspicions toward others (p. 416). But awareness that the owners of those shoes, once identified thanks to those same prints, could have then made a devastating accusation of complicity [chiamata di correità], would certainly have deterred from such an aim. Much better would have been to erase everything. Furthermore, many more bloodstains were found in that bathroom that were not cleaned, as will soon be discussed.

To finish with this matter of a clean-up, contradicted by logical and factual arguments, one must finally consider that no bloodstained cloths were found either on Via Della Pergola or at Sollecito’s apartment (the ruling does not deal at all with this), materials that, according to the decision, would have been bought by Amanda in the early morning of November 2 at Quintavalle’s shop. This witness, however, aside from the reliability of his account, ruled out having seen Ms. Knox making any purchase, much less of that kind – as we have already seen.

Another important circumstance remains unexplained as well: only two traces contain a Meredith – Amanda mixed profile, Exhibit 176 [recte 177] in Romanelli’s room and Exhibit 183 in the corridor. The others can be attributed to Amanda alone, and Exhibit 176, left in Romanelli’s room, even to Meredith alone. If the first-level Court’s explanation were plausible, they should all contain a mixed profile, or, at least, that of only Meredith as well. With a little common sense, however, it is useful to remember that the girls lived together in that house, that they were all friends, and who knows how many times they went up and down barefoot in the various rooms, as often young people do, leaving their traces here and there, even superimposing them by chance, probably on bleach, but without excluding some other possibilities, such as a stain of fruit juice or of vegetable stock.

Indeed – as already recalled above – Dr. Stefanoni (pp. 221 and 228) has acknowledged that it is not possible to date the moment when DNA is released, nor to establish the chronological order in which multiple traces were left, even one above the other. In this respect, then, it must be considered that footprint 183 was collected near [in corrispondenza di] a shoeprint of Guede directed towards the exit, so that this could have been the one to contaminate Amanda’s DNA with Meredith’s. And also concerning contamination, the Police conducted search operations in the rooms of the house before December 18, the day when luminol was employed, and an accidental dragging of genetic material may have occurred.

But the beliefs of the first-level Court in terms of genetics cannot be accepted either. According to Dr. Gino, consultant for the Knox defense, the DNA in those traces is apparently of low quality (low copy number), therefore requiring repetition of the analyses. In the ruling, we read that the credibility and reliability of the analysis derives from the good quality of the instruments, the fact that the latter are subjected to prescribed maintenance, and the correctness of the methodology. It is furthermore verified by the fact that the results are not isolated: the trace with a mixed Meredith – Amanda profile appeared twice, while the one relating to Amanda alone, four times.

In actuality, the quality of the instruments is of no help when the quantity of DNA is insufficient to give reliable results. On this matter, the Conti-Vecchiotti report, commissioned by this Court, has made it clear that below 200 pg, the interpretation of the result is ambiguous and, at any rate, it is necessary to repeat the quantification of the DNA, and also to proceed to multiple confirmatory analyses. In the case of the footprints detected with luminol this healthy and correct practice was not or could not be utilized: this is enough in order not to find any certainty in the results.

To conclude on this subject: the footprints in question, in the opinion of this Court, have no evidentiary [indiziario] value against Amanda Knox.

False Positives and TMB/Luminol Sensitivity

Dr Sarah Gino cited an Australian study of substances that interfere with the forensic luminol test for blood and the nine substances that produce chemiluminescence intensities comparable with that of haemoglobin: turnips, parsnips, horseradishes, commercial bleach [NaClO], copper metal, some furniture polishes, some enamel paints, and some interior fabrics in motor vehicles and based on her own crime scene analysis experience, luminol positive stains are TMB negative for blood 50% of the time.[3] ie, there is a lot of false positives.

This is the study cited by Dr Gino in court: A comprehensive experimental study of industrial, domestic and environmental interferences with the forensic luminol test for blood

TMB is a blood specific presumptive test. In forensic serology, the identification of blood is minimally a two-test procedure and that while confirmatory tests aren’t as sensitive as preliminary ones, they are extremely specific for blood only.[4] Defense expert Adriano Tagliabracci testified TMB is so sensitive it can detect with only 5 blood cells present.[5]

The authors of a review article in Forensic Science Internation on body fluids, Dr. Igor Lednev and Dr. Kelly Virkler, were asked about the luminol-positive, TMB-negative, DNA-negative stains in this case. They responded, “So, there was either no blood and the luminol was wrong, or there was blood and the TMB had interference and the luminol damaged the DNA. We think it is more likely that there was no blood, and that the luminol was reacting with something else, possibly plant matter from the bottom of the shoes causing the footprints (the intensity of the luminol reaction might give some more insight). The prosecution should have used much more convincing evidence to prove the presence of blood.”[6]

In order for a forensic scientist to conclude that a substance is blood, he or she must perform a true confirmatory test. Confirmatory tests detect hemoglobin or glycophorin A, biomolecules found only in blood and they are not prone to the false positive results sometimes obtained with presumptive tests such as luminol and TMB. Previous confirmatory tests such as the Takayama test have been replaced with RSIDTM, which can detect as little as 1 microliter of blood.[7] Another commercial product, HemaTraceTM, has a lower limit of detection of at least one part in 5,000,000.[8] No confirmatory test was ever recorded for the luminol-positive stains.

Besides the negative TMB results and the lack of confirmatory tests, there is one other reason to doubt that the hallway footprints were made in Meredith’s blood. These prints did not give a positive result for anyone’s DNA including Meredith’s. On the one hand a positive result for DNA is not a confirmatory test for blood [other biological tissues have DNA]. Yet on the other hand, a negative result for DNA is evidence against the presence of blood, because white blood cells contain DNA.

The forensic luminol test for blood: unwanted interference and the effect on subsequent analysis by Anders Nilsson, The Swedish National Laboratory of Forensic Science [SKL], Linköping 2006

Webb et al. [22] conducted a study where the luminol test was compared to four other forensic blood tests. These tests where phenolphthalein, LMG, Hemastix [Bayer] and a forensic light source. The luminol test used was found to have by far the greatest sensitivity. Under laboratorial conditions CL was detected from luminol treated stains of the used hemoglobin solution [corresponding to blood] diluted up to 5·106 times. A comparably high sensitivity of the luminol test has been reported in other studies. However the sensitivity is probably not as great under the conditions found at a crime scene and here, depending on several factors, perhaps one may “only” see blood diluted to about 1:10000.

The argument that the TMB reaction was negative because luminol has a lower limit of detection than TMB does is deeply flawed on two grounds. Firstly, there are broad ranges reported for both tests; therefore, it makes sense to compare two values from the same study. For example, Webb, Creamer, and Quickenden report that hemastix [a version of the TMB test] has a lower limit of detection of 1 part in 1,000,000 vs. luminol, which is 1 part n 5,000,000.[9] This is only a fivefold difference, not the 10,000-fold difference calculated elsewhere. Secondly, TMB or another colorimetric test is recommended as a follow-up to a positive luminol result.[10] TMB is not a confirmatory test and positive result has modest value. If every negative result with TMB were solely due to its being less sensitive, there would be no reason to use it whatsoever, one could instead proceed directly to confirmatory testing.

Documents

Description Document Description Document
Hellmann Report: Why the footprints detected with luminol that contain DNA do not prove anything about the murder English Hellmann Report: Why the footprints detected with luminol that do not contain DNA do not prove anything about the murder English
Professor Carlo Torre Italian – English Professor Vinci Luminol Print Report Italian – English
Dr Sarah Gino July 6 Italian Rinaldi-Boemia Footprint Report Italian
Dr Sarah Gino September 26 Italian – English Crime Scene Photos December 18 Wiki GalleryDownload
Patrizia Stefanoni May 22 – Forensics Presentation Italian Crime Scene Video December 18 Part 1Part 2

Resources:

Cottage CSI forgot the luminol, joked about wanting cocaine and didn’t know how to use the equipment UK Daily Mail, October 9, 2011

How Luminol Works

The Luminol Lies by Steve Moore

Mr Hollands Opus by Steve Moore

48 Hours: The Murder of Lanny Horwitz. A case in which the proposition that a positive reaction to luminol proved the presence of blood was advanced by the defence and disputed by the prosecution.

Injustice in Perugia: Sloppy Evidence Collection Procedures and Contamination

The Effect of Luminol on the Serological Analysis of Dried Human blood-stains by Robert Grispino, Special Agent Serology Unit, Laboratory Division FBI Headquarters

The North Carolina Crime Lab Scandal By Paul C. Giannelli. Criminal Justice, Volume 27, Number 1, Spring 2012. © 2012 by the American Bar Association

References:

[1] Stefanoni testimony (p177)

JUDGE – ok! And here there is a degree of sensitivity?
ANSWER – It is very sensitive, now I do not know how to say it to him, however, in common practice …
JUDGE – There also cites false positives of the series …
ANSWER – Yes, in the sense that it does not distinguish whether it is human or animal blood, for example.
JUDGE: However where the result is negative I’m given to understand that it’s almost certain that it is not [blood]?
ANSWER: Yes, it’s not blood, it is not, yes.

[2] Massei Report p256-257

With respect to the Luminol-positive traces found in Romanelli’s room, in Knox’s room and in the corridor, she stated that by analysing the SAL cards “we learn, in contradiction to what was presented in the technical report deposited by the Scientific Police, and also to what was said in Court, that not only was the Luminol test performed on these traces, but also the generic diagnosis for the presence of blood, using tetramethylbenzidine…and this test…gave a negative result on all the items of evidence from which it was possible to obtain a genetic profile”

[3] Massei Report (p258)

“She added that, in her own experience, analyses performed with TMB on traces revealed by Luminol give about even results: 50% negative, 50% positive”

[4] Luminol And The Crime Scene by Robert Grispino, Special Agent Serology Unit, Laboratory Division FBI Headquarters

[5] Tagliabracci testimony (p71)

tetramethylbenzidine is a very sensitive diagnosis that can highlight up to five red blood cells. So that both negative result in short leaves no room for doubt…

[6] Forensic tests for the presence of blood by Professor Chris Halkides

[7] Independent Forensics Rapid Stain Identification Of Human Blood (RSID™-Blood) Alexander Sinelnikov *, Anna Kalinina, Jennifer B. Old, Pravatchai W. Boonlayangoor
and Karl A. Reich

[8] Canadian Society of Forensic Science Volume 36, Number 3. September 2003

[9] Chemiluminescence in the Visualization of Forensic Bloodstains (p2)

[10] Webb JL, Creamer JI, Quickenden TI. Luminescence 2006; 21: pp. 214–220, DOI: 10.1002/bio.908, “A comparison of the presumptive luminol test for blood with four non-chemiluminescent forensic techniques” abstract: http://www.ncbi.nlm.nih.gov/pubmed/16645959

[11] Gefrides, LA, and Welch KE. “Serology and DNA” in The Forensic Laboratory Handbook Procedures and Practice, Humana Press, 2006.

Arnone-Ippolito-Transcript (OCR) page 84

PM Comodi: Now I wanted to clarify the following. The two gentlemen that we have called today, the first being Arnone and just now Mr. Ippolito, are those who actually took these videos and the photos of the second site visit, they did not carry out any analysis, because the collection of samples, evidence collection, etc, wasn’t done by them, they only recorded it.

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